Rôle de la cytokine inflammatoire IL-15 dans la progression de la fibrose hépatique

Abstract: Chronic inflammation in the liver causes liver fibrosis, which predisposes to liver cancer. Indeed, most cases of liver cancer occur in fibrotic livers. Different immune cells specifically NK cells and macrophages and various cytokines and chemokines contribute to the progression of fibrosis as well as its resolution. Interleukin-15 (IL-15) is a pro-inflammatory cytokine produced mainly by immune cells in response to innate immune stimuli and is required for immune cell infiltration and homeostasis in the liver micromilieu. However, the exact role of IL-15 in the homeostasis of liver microenvironment remains to be elucidated. Hence, the goal of this thesis is to determine the role of IL-15 in the progression and pathogenesis of liver fibrosis. Liver fibrosis was experimentally induced in wildtype (WT) and Il15-/- mice either by intraperitoneal injection of carbon tetrachloride (CCl4) twice a week for 7 weeks or by providing choline-deficient high-fat-diet (CD-HFD) ad libitum for 12 weeks. After the induction period, mice were sacrificed, and liver tissues were collected for histology (H&E and Sirius Red), immunohistochemistry (α-SMA and Collagen) and immunofluorescence (CD68). Proinflammatory cytokine gene expression was assessed by qRT-PCR. In WT mice, CCl4 treatment significantly increased IL-15 expression in the liver along with increased collagen deposition and elevated proportion of α-smooth muscle actin (α-SMA) positive cells in liver sections. Similar observations were made in CD-HFD induced liver fibrosis model also. Both collagen deposition and α-SMA positive cells were significantly reduced in the livers of CCl4-treated or CD-HFD fed IL-15 deficient mice. Concomitantly, the induction of various proinflammatory cytokines (Il12, Tnfa, Tgfb and iNos) and chemokines (Ccl2, Ccl5, Cxcl9 and Cxcl10) following CCl4 treatment was significantly reduced in Il15-/- mice compared to control mice. Additionally, the livers of CD-HFD fed WT mice also showed reduced Ppara expression indicating that IL-15 mediated inflammation alters lipid metabolism. Our results suggest that IL-15 mediated inflammatory response may have an important role in the progression of liver fibrosis. Our data indicate a pro-fibrogenic role of IL-15 in the pathogenesis of liver fibrosis and raises the possibility of targeting IL-15 signaling for the management of for liver fibrosis.L'inflammation chronique du foie provoque une fibrose, qui prédispose au cancer du foie. En effet, la plupart des cas de ces cancer surviennent dans les cas fibrotiques. Différentes cellules immunitaires, en particulier les cellules NK et les macrophages ainsi que diverses cytokines et chimiokines contribuent à la progression de la fibrose et à sa résolution. L'interleukine-15 (IL-15) est une cytokine pro-inflammatoire produite principalement par les cellules immunitaires en réponse à des stimuli immunitaires innés. Elle est nécessaire pour l'infiltration de cellules immunitaires dans le foie. Cependant, le rôle exact de l'IL-15 dans l'homéostasie du microenvironnement du foie reste à élucider. Par conséquent, l'objectif de cette thèse est de déterminer le rôle de l'IL-15 dans la progression et la pathogenèse de la fibrose du foie. La fibrose hépatique a été induite expérimentalement soit par injection intrapéritonéale de tétrachlorure de carbone (CCl4), soit en fournissant un régime riche en graisses et déficient en choline (CD-HFD) à des souris du type sauvage (WT) ou Il15-/-. Le CCl4 a été administré par voie intrapéritonéale deux fois par semaine pendant 7 semaines et les souris ont un accès illimité à la nourriture CD-HFD pendant 12 semaines. Après la période d'induction, les souris ont été sacrifiées et les tissus du foie ont été recueillis pour l'histologie (H&E et Sirius Red), l'immunohistochimie (α-SMA et le collagène) et l'immunofluorescence (CD68). L'expression du gène de la cytokine pro-inflammatoire a été évaluée par qRT-PCR. Chez les souris WT, le traitement par CCl4 a augmenté de manière significative l’expression de l’IL-15 dans le foie, ainsi que le dépôt de collagène et une proportion élevée de cellules positives pour l’actine du muscle lisse (α-SMA) dans les coupes de foie. Des observations similaires ont été observées dans le modèle de fibrose hépatique induite par CD-HFD. Les dépôts de collagène et les cellules positives pour SMA ont été significativement réduits dans le foie des souris déficientes en IL-15 traitées avec CCl4 ou traitées avec CD-HFD. Parallèlement, l'induction de diverses cytokines pro-inflammatoires (Il12, Tnfa, Tgfb et iNos) et de chimiokines (Ccl2, Ccl5, Cxcl9 et Cxcl10) après traitement par CCl4 a été significativement réduite chez la souris Il15-/- par rapport à la souris contrôle. De plus, les foies des souris WT nourries avec CD-HFD ont également montré une expression réduite du gène Ppara, ce qui indique que l'inflammation médiée par l'IL-15 altère le métabolisme des lipides. Nos résultats suggèrent que la réponse inflammatoire médiée par l'IL-15 pourrait jouer un rôle important dans la progression de la fibrose hépatique. Nos données indiquent un rôle pro-fibrogénique de l'IL-15 dans la pathogenèse de la fibrose hépatique et soulèvent la possibilité de cibler la signalisation de l'IL-15 pour le traitement de la fibrose hépatique

ACUTE TOXICITY EVALUATION OF PROTODIOSCIN RICH EXTRACT OF TRIGONELLA FOENUM-GRAECUM L IN RATS

ABSTRACTObjectives: The objective of this study was to investigate the acute toxicity of standardized protodioscin rich extract (PRE) of Trigonella foenumgraecumL (26.63% w/w; PRE).Methods: To evaluate the toxicity of PRE, the acute toxicity of the extract on adult rats were investigated. A fixed large dose of 2 g/kg body weight ofPRE was administrated by a single oral gavage, and 1 g/kg body weight of PRE was administered by intravenous according to the Organisation forEconomic Co-operation and Development guidelines.Results: In 2 weeks, PRE showed no obvious acute toxicity. There were no deaths in either group and no change in the clinical signs. The hematologicaland biochemical analyses showed some changes that returned to reference levels without impairment of homeostasis. The treatment did not induceuntoward changes in organs as shown by histological studies. The in vivo results showed that has low toxicity.Conclusion: PRE is safe and can be potentially used as an aphrodisiac in future studies.Keywords: Protodioscin, Aphrodisiac, Trigonella foenum-graecum, Organisation for Economic Co-operation and Development, Toxicity,Histopathological studies

A LITERATURE REVIEW: THE ROLE OF KSHARA IN AYURVEDA

Ayurveda depends largely upon the medicinal plants for the therapy. Among the four Vedas - Rig-Veda, Yajurveda, Samaveda, and Atharvaveda - the Atharvaveda is considered to be one from which Ayurveda is derived and several medicinal plants are mentioned that can be used as a drugs. Kshara is the herbal extracts of plants, According to Sushruta, in Kshara Paka Vidhi Adhyaya has mentioned 23 drugs plants as: Mushkka, kutaj, Palash, Ashwakarna, Paribhadrak, Bhibitaka, Aragvadha, Tilwaka, Arka, Snuhi, Apamarga, Patla, Naktamal, Vrusha, Kadali, Chitraka, Putika, Asphota, Ashwamarak, Saptachanda, Agnimantha, Gunja, Koshataki from which Kshara can be prepared. Kshara can be a multiple combination of many herbs or may be from single herb. Kshara is useful in internal medicines in different formulation. i.e., Kshara Gutika (tablet), Vati, Ksharavleha, Kshara-Ghrita. Kshara is also useful in external application directly as Ksharapichu (gause-piece), Kshara Sutra, Ksharavarti. This alkaline preparation has many therapeutic usages and many surgical procedures. Ksharakarma is useful as the substitute of surgical instruments, because they can be used safely on the patients who are afraid of surgery. It has been proved to be effective in treating many disorders like Dushta Vrana (wounds), Bhagandara (Fistula-in-ano), Arsha (haemorrhoids), Charmakeela, Ashmari (Stone), Gulma Udara (ascites), Kushtha (leprosy), Rohini etc. External application of Kshara is indicated in children, weak persons and decrepit. Ksharakarma has been considered as a wealth and a strong weapon in Ayurvedic Pharmacopeia. In this paper we have described Preparation, classification, properties, Guna, Dosha, Indication, contra-indication, different formulae used in medicine, Matra, chemical composition and assessment.

IN SILICO MOLECULAR DOCKING AND PHARMACOKINETIC PREDICTION OF GALLIC ACID DERIVATIVES AS PPAR-γ AGONISTS

Objective: To perform molecular docking and pharmacokinetic prediction of gallic acid derivatives as Peroxisome proliferator-activated receptors-γ (PPAR-γ) agonist for the treatment of diabetes.Methods: Molecular docking study on gallic acid and different derivatives of gallic acid was performed using GOLD v5.2 software. In addition to this, all the derivatives were analysed for drug likeliness, Lipinski's rule and ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties using online tools like admet SAR, Molinspiration and Medchem designer.Results: Molecular docking studies reveals that SSP-12, SSP-13 and SSP-40 demonstrated significant binding to the PPAR-γ receptor with good Gold score fitness (73.11, 69.86 and 75.51 respectively) and relative ligand energy (-8.26,-8.33 and-7.82, respectively) as compared to standard drugs i.e. rosiglitazone and pioglitazone, (64.10 and 66.72) and (-4.30 and-2.47) respectively.Conclusion: The final results of molecular docking along with information gathered from pharmacokinetic parameters of gallic acid derivatives may be utilised further for the development of newer PPAR-γ agonists having anti-diabetic potential with better pharmacokinetic and pharmacodynamic profile

Synthesis, anti-tuberculosis and anti-bacterial activities of sulfonamide bearing 4-((2-(5-bromo-1H-pyrazolo[3,4-b]pyridin-1-yl)-2-oxoethyl)amino)-N-(various substitutions)benzenesulfonamide

682-689In this study all the new synthesized compounds of sulfonamide bearing 4-((2-(5-bromo-1H-pyrazolo[3,4-b]pyridin-1-yl)-2-oxoethyl)amino)-N-(various-substitutions)benzenesul-fonamide have been synthesized by using 1-(5-bromo-1H-pyarazolo[3,4-b]pyridin-1yl)-2-chloroethanone which has been fused with various sulfa drugs in presence of potassium carbonate and DMF. All the derivatives have been recognized by physical properties like melting point and characterization by elemental analysis (CHNS) and various spectral techniques such as FT-IR, 1H and 13C NMR, and ESI-MS. The series of these pyrazolo[3,4-b]pyridin bearing sulfonamide have been synthesized and the final derivatives have been evaluated for bioactivity such as anti-bacterial activity against gram +ve and gram −ve and also screened for their in vitro anti tubercular activity against Mycobacterium tuberculosis H37RV. The results for the synthesized compounds have been compared against standard drugs

Synthesis and characterization of 4(3-(4-Fluorophenyl)2-methyl-1-(4-(sulfonamidesubtituted)phenyl)-4-oxoazitidin-2-yl)-3-methyl-1-(p-tolyl)-1H-pyrazol-5(4H)One as Antibacterial, and Antioxidant Candidates

A series of all novels 4(3-(4-Fluorophenyl)2-methyl-1-(4-(sulfonamidesubtituted)phenyl)-4-oxoazitidin-2-yl)-3-methyl-1-(p-tolyl)-1H-pyrazol-5(4H) One 5a-5r poly functionalized derivatives were containing sulfonamide functionality united with 2-Azitidinone (Azitidin-2-one or β-lactam) group which designed and synthesized with moderate to good yield. The starting with 4-acyl-2-pyrazolin-5-one (APYZ) 1 which condensed with different sulfonamides 2 produced intermediate Schiff bases 4-(arylideneamino)-N-(thiazol-2-yl)benzensulfonamide 3a-r were cyclization with 2-(4-flourophenyl)chloroacetylchloride (F-CAC) 4 which produced targeted compounds 5a-5r of 2-Azitidinone versatile group in good yield. The isolated compounds were recognized by spectral and elemental investigation. The compounds 5f, 5l, 5n, and 5r showed excellent increased antibacterial activity compared to streptomycin standard drug and 5c, 5f, 5l, 5o, and 5r showed moderate to good antioxidant properties with used DPPH radical scavenging assay

Predicting optical coherence tomography-derived diabetic macular edema grades from fundus photographs using deep learning

Diabetic eye disease is one of the fastest growing causes of preventable blindness. With the advent of anti-VEGF (vascular endothelial growth factor) therapies, it has become increasingly important to detect center-involved diabetic macular edema (ci-DME). However, center-involved diabetic macular edema is diagnosed using optical coherence tomography (OCT), which is not generally available at screening sites because of cost and workflow constraints. Instead, screening programs rely on the detection of hard exudates in color fundus photographs as a proxy for DME, often resulting in high false positive or false negative calls. To improve the accuracy of DME screening, we trained a deep learning model to use color fundus photographs to predict ci-DME. Our model had an ROC-AUC of 0.89 (95% CI: 0.87-0.91), which corresponds to a sensitivity of 85% at a specificity of 80%. In comparison, three retinal specialists had similar sensitivities (82-85%), but only half the specificity (45-50%, p<0.001 for each comparison with model). The positive predictive value (PPV) of the model was 61% (95% CI: 56-66%), approximately double the 36-38% by the retinal specialists. In addition to predicting ci-DME, our model was able to detect the presence of intraretinal fluid with an AUC of 0.81 (95% CI: 0.81-0.86) and subretinal fluid with an AUC of 0.88 (95% CI: 0.85-0.91). The ability of deep learning algorithms to make clinically relevant predictions that generally require sophisticated 3D-imaging equipment from simple 2D images has broad relevance to many other applications in medical imaging

Role of β-Interferon Inducer (DEAE-Dextran) in Tumorigenesis by VEGF and NOTCH1 Inhibition along with Apoptosis Induction

As a novel target for breast cancer, interferon inducers have found its role as anti-angiogenic agents with diethylaminoethyl dextran (DEAE-Dextran) being a molecule used for centuries as a transfection agent. Our results herein offer an explanation for the emergence of DEAE-Dextran as an anti-tumor agent for TNBC with in-depth mechanistic approach as an anti-angiogenic molecule. DEAE-Dextran has found to possess cytotoxic activity demonstrated during the various in vitro cytotoxicity assays; moreover, as an anti-oxidant, DEAE-Dextran has shown to possess excellent reactive oxygen species scavenging activity. The interferon inducing capacity of DEAE-Dextran was determined qualitatively as well as quantitatively specifically demonstrating overexpression of β-interferon. As a measure of anti-proliferative activity, DEAE-Dextran exhibited reduced ki67, p53, and PCNA levels. Also, overexpression of CK5/6 and p63 in DEAE-Dextran treated animals indicated improvement in breast cell morphology along with an improvement in cell–cell adhesion by virtue of upregulation of β-catenin and E-cadherin. Anti-angiogenic property of DEAE-Dextran was concluded by the downregulation of CD31, VEGF, and NOTCH1 both in vivo and in vitro. Further, apoptosis due to DEAE-Dextran, initially determined by downregulation of Bcl2, was confirmed with flow cytometry. Overall, results are defensive of DEAE-Dextran as an emerging anti-tumor agent with mechanisms pertaining to β-interferon induction with probable VEGF and NOTCH1 inhibition as well as apoptosis which still needs to be studied in further depth